BLT-PF usage FAQ for Illumina DNA PCR-Free

Note:The BLT-PF reagent is the bead linked transposome reagent used in the Illumina DNA PCR-Free workflow.

Can 10 µl of BLT-PF be used (like in the integrated extraction protocol) for purified gDNA? No, this will impact sequencing metrics.

If saliva/blood input is quantified, can 15 µl BLT-PF be used instead? What would happen (in the Tagment Genomic DNA section of Reference Guide)? This will increase the amount of DNA required for the integrated normalization. For some samples, this may negatively impact the self-normalization feature of the Illumina DNA PCR-Free method.

Why is there a different amount of BLT-PF if using gDNA vs blood or saliva input? This is intended to make sure that raw samples are normalized as they are processed through the workflow, allowing for utilization of the normalizing effect of the bead linked transposome.

Should dried blood spots (DBS) be treated like gDNA (using 15 µl BLT-PF) or like blood/saliva (using 10 µl BLT-PF) in the Tagment Genomic DNA section of the Reference Guide? Treat dried blood spots like blood/saliva samples, using 10 µl BLT-PF for each sample.

What happens if the beads settle out of solution?

Resuspend the beads and proceed as normal.

For any feedback or questions regarding this article (Illumina Knowledge Article #3453), contact Illumina Technical Support [e-mail].